Background:Chortoglyphus arcuatus has been described in many countries. Many allergens are potent enzymes, which may promote a Th2 immune response. The aim of this study was to evaluate the enzymatic activity of body and fecal extracts of C. arcuatus. Material and Methods: Feces and bodies of full-grown C. arcuatus cultures were separated by sieving, extracted in PBS, dialyzed and lyophilized. The antigenic profile of both extracts was determined by SDS-PAGE. Immunoblot experiments were conducted using a pool of sera from allergic individuals residing in Galicia, a region of Spain, where this species is abundant. The enzymatic activity of the extracts was evaluated by the zymogram technique. Serine and cysteine protease activity was measured using in vitro methods. The API Zym® system was used to determine the enzymatic properties of the extracts. Results: The antigenic profile showed that the body extract contained more and better defined bands than the fecal extract. Allergens were detected in both extracts in a molecular weight range between 14 and 100 kDa. Gelatinolytic gels confirmed that fecal extracts contain more hydrolytic enzymatic activity than body extracts. Serine protease activity in fecal extracts was higher than in body extracts (5.98 vs. 2.701 IU of trypsin/mg of freeze-dried material). No cysteine protease activity was detected. Conclusion:C. arcuatus extracts contain several allergens and proteins with high enzymatic activity, especially in the feces. Some of these allergens may be enzymes. Fecal extracts have more enzymatic activity than body extracts.

Keywords:
Chortoglyphus arcuatus, Enzymatic activity, allergen, Mites
1.
Boquete M, Iraola V, Fernández-Caldas E, Arenas Villaroel L, Carballada FJ, Gonzalez de la Cuesta C, Lopez-Rico MR, Nuñez Orjales R, Parra GA, Soto-Mera MT, Varela S, Vidal C: House dust mite species and allergen levels in Galicia, Spain: a cross-sectional, multicenter, comparative study. J Investig Allergol Clin Immunol 2006;16:169–176.
2.
Boquete M, Carballás C, Carballada F, Iraola V, Carnés J, Fernández-Caldas E: In vivo and in vitro allergenicity of the domestic mite Chortoglyphus arcuatus. Ann Allergy Asthma Immunol 2006;97:203–208.
3.
Fernández-Caldas E, Vailes LD, Troubina O, King EM, Arlian L, Boquete M, Iraola V, Lafosse-Marin S, Chapman MD: Molecular cloning of Chortoglyphus arcuatus allergens. J Allergy Clin Immunol, in preparation.
4.
Marx JJ Jr, Twiggs JT, Ault BJ, Merchant JA, Fernández-Caldas E: Inhaled aeroallergen and storage mite reactivity in a Wisconsin farmer nested case-control study. Am Rev Respir Dis 1993;147:354–358.
5.
Yadav A, Elder BL, Morgan MS, Vyszenski-Moher DL, Arlian LG: Prevalence of serum IgE to storage mites in a southwestern Ohio population. Ann Allergy Asthma Immunol 2006;96:356–362.
6.
Puerta L, Fernandez-Caldas E, Lockey RF, Caraballo LR: Sensitization to Chortoglyphus arcuatus and Aleuroglyphus ovatus in Dermatophagoides spp. allergic individuals. Clin Exp Allergy 1993;23:117–123.
7.
Morgan MS, Arlian LG: Enzymatic activity in extracts of allergy-causing astigmatid mites. J Med Entomol 2006;43:1200–1207.
8.
Fernández-Caldas E, Iraola Calvo V: Mite allergens. Curr Allergy Asthma Rep 2005;5:402–410.
9.
Bowman CE: Comparative enzymology of economically important astigmatid mites; in Griffiths DA, Bowman CE (eds): Acarology VI. Chichester, Ellis-Horwood, 1984, vol 2, p 993.
10.
Sánchez-Ramos I, Hernandez CA, Castanera P, Ortego F: Proteolytic activities in body and faecal extracts of the storage mite, Acarus farris. Med Vet Entomol 2004;18:378–386.
11.
Ortego F, Sanchez-Ramos I, Ruiz M, Castanera P: Characterization of proteases from a stored product mite, Tyrophagus putrescentiae. Arch Insect Biochem Physiol 2000;43:116–124.
12.
Montealegre F, Quinones C, Torres N, Goth K: Detection of serine proteases in extracts of the domestic mite Blomia tropicalis. Exp Appl Acarol 2002;26:87–100.
13.
Flores I, Mora C, Rivera E, Donnelly R, Montealegre F: Cloning and molecular characterization of a cDNA from Blomia tropicalis homologous to dust mite group 3 allergens (trypsin-like proteases). Int Arch Allergy Immunol 2003;130:12–16.
14.
Cardona G, Guisantes J, Eraso E, Serna LA, Martinez J: Enzymatic analysis of Blomia tropicalis and Blomia kulagini (Acari: Echimyopodidae) allergenic extracts obtained from different phases of culture growth. Exp Appl Acarol 2006;39:281–288.
15.
Nisbet AJ, Billingsley PF: Hydrolytic enzymes of Psoroptes cuniculi (Delafond). Insect Biochem Mol Biol 1999;29:25–32.
16.
Edwards TB, Trudeau WL, Fernández-Caldas E, Lee DK, Seleznick MJ, Lockey RF: Proteinases in extracts of the storage mite, Aleuroglyphus ovatus. J Allergy Clin Immunol 1992;90:129–131.
17.
Ando T, Homma R, Ino Y, Ito G, Miyahara A, Yanagihara T, Kimura H, Ikeda S, Yamakawa H, Iwaki M, et al: Trypsin-like protease of mites: purification and characterization of trypsin-like protease from mite faecal extract Dermatophagoides farinae. Relationship between trypsin-like protease and Der f III. Clin Exp Allergy 1993;23:777–784.
18.
Stewart GA, Lake FR, Thompson PJ: Faecally derived hydrolytic enzymes from Dermatophagoides pteronyssinus: physicochemical characterisation of potential allergens. Int Arch Allergy Appl Immunol 1991;95:248–256.
19.
Kikuchi Y, Takai T, Kuhara T, Ota M, Kato T, Hatanaka H, Ichikawa S, Tokura T, Akiba H, Mitsuishi K, Ikeda S, Okumura K, Ogawa H: Crucial commitment of proteolytic activity of a purified recombinant major house dust mite allergen Der p 1 to sensitization toward IgE and IgG responses. J Immunol 2006;177:1609–1617.
20.
Michaud D, Faye L, Yelle S: Electrophoretic analysis of plant cysteine and serine proteinases using gelatin-containing polyacrylamide gels and class-specific proteinase inhibitors. Electrophoresis 1993;14:94–98.
21.
Weghofer M, Thomas WR, Pittner G, Horak F, Valenta R, Vrtala S: Comparison of purified Dermatophagoides pteronyssinus allergens and extract by two-dimensional immunoblotting and quantitative immunoglobulin E inhibitions. Clin Exp Allergy 2005;35:1384–1391.
22.
Pittner G, Vrtala S, Thomas WR, Weghofer M, Kundi M, Horak F, Kraft D, Valenta R: Component-resolved diagnosis of house-dust mite allergy with purified natural and recombinant mite allergens. Clin Exp Allergy 2004;34:597–603.
23.
Sehgal N, Custovic A, Woodcock A: Potential roles in rhinitis for protease and other enzymatic activities of allergens. Curr Allergy Asthma Rep 2005;5:221–226.
24.
Brown A, Farmer K, MacDonald L, Kalsheker N, Pritchard D, Haslett C, Lamb J, Sallenave JM: House dust mite Der p 1 downregulates defenses of the lung by inactivating elastase inhibitors. Am J Respir Cell Mol Biol 2003;29(3 Pt 1):381–389.
25.
Custovic A, Simpson A, Woodcock A: Importance of indoor allergens in the induction of allergy and elicitation of allergic disease. Allergy 1998;53:115–120.
© 2007 S. Karger AG, Basel
2007
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.